Daan Pieren

91 Compromised DNA repair promotes the accumulation of regulatory T cells A B C Ercc1 +/+ (4 months) Ercc1 -/ Δ 7 (4 months) 70 80 90 100 %ThofnaiveCD4 + ** 0 2 4 6 8 10 % CD25 + ofTh ** 0 2 4 6 8 10 % CD122 + ofTh ns 0 2 4 6 8 10 % PD-1 + ofTh ns 0 20 40 60 80 100 %ThofmemoryCD4 + ns 0 5 10 15 % CD25 + ofTh ns 0 20 40 60 80 100 % CD122 + ofTh *** 0 10 20 30 40 % PD-1 + ofTh 0.09 0 10 20 30 %TregofnaiveCD4 + ** 70 80 90 100 % CD25 + ofTreg ** 0 2 4 6 8 10 % CD122 + ofTreg ns 0 10 20 30 % PD-1 + ofTreg * 0 20 40 60 %TregofmemoryCD4 + ns 40 60 80 100 % CD25 + ofTreg ns 0 2 4 6 8 10 % CD122 + ofTreg ns 0 20 40 60 80 % PD-1 + ofTreg ns 0 20 40 60 %naiveTcofCD3 + ** 0 2 4 6 8 10 %memoryTcofCD3 + * 0 2 4 6 8 10 % CD25 + ofTc ns 0 5 10 15 20 % CD25 + ofTc ns 0 2 4 6 8 10 % CD122 + ofTc ns 0 20 40 60 80 100 % CD122 + ofTc ** 0 2 4 6 8 10 % PD-1 + ofTc * 0 10 20 30 % PD-1 + ofTc ** 0 2 4 6 8 10 CD25MFIofTh ns 0 50 100 150 CD25MFIofTh ** 0 1 2 3 4 5 CD122MFIofTh ns 0 50 100 150 CD122MFIofTh ** 0 50 100 150 PD-1MFIofTh ns 0 100 200 300 400 PD-1MFIofTh ** 0 1000 2000 3000 CD25MFIofTreg ** 0 500 1000 1500 2000 CD25MFIofTreg ns 0 50 100 150 CD122MFIofTreg ns 0 20 40 60 80 100 CD122MFIofTreg ns 0 50 100 150 200 250 PD-1MFIofTreg * 0 200 400 600 800 PD-1MFIofTreg ns 0 2 4 6 8 10 CD25MFIofTc ns 0 10 20 30 CD25MFIofTc ns 0 20 40 60 80 100 CD122MFIofTc ns 0 100 200 300 400 500 CD122MFIofTc ** 0 5 10 15 20 25 PD-1MFIofTc ns 0 20 40 60 80 PD-1MFIofTc ** Naive Th Memory Th Naive Treg Memory Treg Naive Tc Memory Tc T-cell subsets %CD25 + %CD122 + %PD-1 + CD25MFI CD122 MFI PD-1 MFI Supplementary Figure 4. CD25 + , CD122 + , and PD-1 + cell frequencies and expression within naive and memory T-cell subsets. Bar graphs ( A ) indicate the frequencies of the following T-cell subsets: naive Th cells (FoxP3 - cells of CD44 Lo CD4 + CD3 + ), memory Th cells (FoxP3 - cells of CD44 Hi CD4 + CD3 + ), naive Treg cells (FoxP3 + cells of CD44 Lo CD4 + CD3 + ), memory Treg cells (FoxP3 + cells of CD44 Hi CD4 + CD3 + ), naive Tc cells (CD44 Lo CD4 - of CD3 + ), and memory Tc cells (CD44 Hi CD4 - of CD3 + ). Within each of these T-cell subsets ( B ) the frequencies of CD25 + , CD122 + , and PD-1 + cells were determined, as well as ( C ) the median expression intensity (MFI) of CD25, CD122, and PD-1. Bar graphs show mean ± SD; * p < 0.05, ** p < 0.01, *** p < 0.001, ns = not statistically significant for the difference between groups using parametric unpaired Student’s t test or non-parametric Mann-Whitney test, two-tailed, dependent on the tested normality of distribution of the data. 3